
PROTOCOLS
Extraction of RNA from Lipid Tissues
Paul Zumbo
Protocol for the extraction of RNA from lipid tissues (≤ 10mg). This process disrupts lipid tissues with a pellet pestle in the presence of a mono-phasic solution containing chaotropic denaturants phenol and guanidine thiocyanate. The solution is homogenized using a syringe and 21 g needle, and separated into an aqueous phase and organic phase with the addition of chloroform. The RNA is recovered from the aqueous phase and purified using silica-based membrane technology with on column DNase digestion. RNA molecules < 200 nt are selectively excluded.
Isolation of Total RNA from Animal Cells
Paul Zumbo
Protocol for the extraction of RNA from < 5×10⁵ animal cells. This process harvests and disrupts human or animal cells in the presence of a mono-phasic solution containing chaotropic denaturants phenol and guanidine thiocyanate. The solution is homogenized using a syringe and 21 g needle, and separated into an aqueous phase and organic phase with the addition of chloroform. The RNA is recovered from the aqueous phase and purified using silica-based membrane technology with on column DNase digestion. RNA molecules < 200 nt are selectively excluded.
Purification of Total RNA with DNase Treatment
Paul Zumbo
Protocol for purifying total RNA with DNase treatment. This process purifies and concentrates RNA (< 45 ug) and removes contaminant DNA using silica-based membrane technology and DNase digestion. All RNA molecules < 200 nt are selectively excluded.

